A Narcissus mosaic viral vector system for protein expression and flavonoid production.
A Narcissus mosaic viral vector system for protein expression and flavonoid production.
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DOI：
10.1186/1746-4811-9-28
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发表时间：
2013-07-13
期刊：
影响因子：
5.1
通讯作者：
Davies K
中科院分区：
文献类型：
--
作者：
Zhang H;Wang L;Hunter D;Voogd C;Joyce N;Davies K
随着下一代测序产生的序列数量的爆炸性增长，对高通量筛选以了解基因功能的需求也在增长。植物病毒载体已被广泛用作下调植物基因表达的工具。然而，植物病毒载体也可以以非常有效的方式表达蛋白质，因此也可以作为表征蛋白质及其在植物代谢途径中的功能的有价值的工具。在这项研究中，我们开发了一个基于Gateway®的高通量水仙花叶病毒（NMV）病毒载体克隆系统。利用GFP和GUS报告基因、植物基因PAP 1（一个激活花青素途径的R2 R3 MYB）和硒结合蛋白1（SeBP），我们证明了NMV载体和模式植物本氏烟草（Nicotianabenthamiana）可用于高效的蛋白表达、蛋白亚细胞定位和次生代谢产物的产生。我们的研究结果表明，植物病毒载体系统不仅可以用于蛋白质的工作，但也可以潜在地适合于生产有价值的次生代谢产物在一个大规模的，因为该系统不需要从种子或愈伤组织，这是某些次生代谢产物可以干扰发展阶段的植物再生。
With the explosive numbers of sequences generated by next generation sequencing, the demand for high throughput screening to understand gene function has grown. Plant viral vectors have been widely used as tools in down-regulating plant gene expression. However, plant viral vectors can also express proteins in a very efficient manner and, therefore, can also serve as a valuable tool for characterizing proteins and their functions in metabolic pathways in planta. In this study, we have developed a Gateway®-based high throughput viral vector cloning system from Narcissus Mosaic Virus (NMV). Using the reporter genes of GFP and GUS, and the plant genes PAP1 (an R2R3 MYB which activates the anthocyanin pathway) and selenium-binding protein 1 (SeBP), we show that NMV vectors and the model plant Nicotiana benthamiana can be used for efficient protein expression, protein subcellular localization and secondary metabolite production. Our results suggest that not only can the plant viral vector system be employed for protein work but also can potentially be amenable to producing valuable secondary metabolites on a large scale, as the system does not require plant regeneration from seed or calli, which are stages where certain secondary metabolites can interfere with development.
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影响因子：
3
作者：
Albert, Nick W.;Arathoon, Steve;Davies, Kevin M.
通讯作者：
Davies, Kevin M.
影响因子：
4.8
作者：
Hunter DA;Fletcher JD;Davies KM;Zhang H
通讯作者：
Zhang H
影响因子：
7.2
作者：
BAULCOMBE, DC;CHAPMAN, S;CRUZ, SS
通讯作者：
CRUZ, SS
影响因子：
7.2
作者：
Albert, Nick W.;Lewis, David H.;Davies, Kevin M.
通讯作者：
Davies, Kevin M.
影响因子：
13.8
作者：
Fukuzawa, Noriho;Ishihara, Takeaki;Matsumura, Takeshi
通讯作者：
Matsumura, Takeshi